From near extinction to diversification by means of a shift in pollination mechanism in the gymnosperm relict Ephedra (Ephedraceae,Gnetales)

Pollination in gymnosperms is usually accomplished by means of wind, but some groups are insect‐pollinated. We show that wind and insect pollination occur in the morphologically uniform genus Ephedra (Gnetales). Based on field experiments over several years, we demonstrate distinct differences between two Ephedra species that grow in sympatry in Greece in pollen dispersal and clump formation, insect visitations and embryo formation when insects are denied access to cones. Ephedra distachya, nested in the core clade of Ephedra, is anemophilous, which is probably the prevailing state in Ephedra. Ephedra foeminea, sister to the remaining species of the genus, is entomophilous and pollinated by a range of diurnal and nocturnal insects. The generalist entomophilous system of E. foeminea, with distinct but infrequent insect visitations, is in many respects similar to that reported for Gnetum and Welwitschia and appears ancestral in Gnetales. The Ephedra lineage is well documented already from the Early Cretaceous, but the diversity declined dramatically during the Late Cretaceous, possibly to near extinction around the Cretaceous–Palaeogene boundary. The clade imbalance between insect‐ and wind‐pollinated lineages is larger than expected by chance and the shift in pollination mode may explain why Ephedra escaped extinction and began to diversify again.     

Beating the blaze: Fire survival in the fan aloe (Kumara plicatilis), a succulent monocotyledonous tree endemic to the Cape fynbos,South Africa

Fire is central to the ecology of Mediterranean‐type climate ecosystems, but little is known about the fire ecology of succulent plants therein. This study investigated the fire ecology of an arborescent succulent monocot, Kumara plicatilis (L.) G. D. Rowley (Asphodelaceae), a Cape fynbos endemic. Habitat suitability was assessed to determine whether the species tolerates or ‘avoids’ fire, and fire survival traits (bark thickness and tissue water content) were measured. The population size structure and density of three K. plicatilis populations were assessed after natural fires, and resprouting potential was investigated. Kumara plicatilis adopts a dual fire survival strategy, occupying rocky sites to ‘avoid’ fire and possessing morphological features that afford fire tolerance, e.g. well‐protected apical meristems and thick corky bark. Bark thickness of burned individuals in situ was similar to unburned plants, suggesting that K. plicatilis bark provides effective insulation against fire. Mortality rates were 64%, 40% and 11%, and decreased as rock cover at the population level increased. All three populations showed reduced plant density post‐fire, with greater density reductions associated with lower rock cover. Small plants appear most vulnerable to fire damage due to lower absolute bark thickness and plant heights within the flame zone. Kumara plicatilis is an apical sprouter, recovering after fire or mechanical stem damage by onward growth from surviving stem apices, rather than resprouting. Post‐fire population recovery therefore likely depends on inter‐fire recruitment.     

Geographic variation in seed traits within and among forty‐two species of Rhododendron (Ericaceae) on the Tibetan plateau: relationships with altitude,habitat, plant height,and phylogeny

Seed mass and morphology are plant life history traits that influence seed dispersal ability, seeding establishment success, and population distribution pattern. Southeastern Tibet is a diversity center for Rhododendron species, which are distributed from a few hundred meters to 5500 m above sea level. We examined intra‐ and interspecific variation in seed mass and morphology in relation to altitude, habitat, plant height, and phylogeny. Seed mass decreased significantly with the increasing altitude and increased significantly with increasing plant height among populations of the same species. Seed mass differed significantly among species and subsections, but not among sections and subgenera. Seed length, width, surface area, and wing length were significantly negative correlated with altitude and significantly positive correlated with plant height. Further, these traits differed significantly among habitats and varied among species and subsection, but not among sections and subgenera. Species at low elevation had larger seeds with larger wings, and seeds became smaller and the wings of seeds tended to be smaller with the increasing altitude. Morphology of the seed varied from flat round to long cylindrical with increasing altitude. We suggest that seed mass and morphology have evolved as a result of both long‐term adaptation and constraints of the taxonomic group over their long evolutionary history.     

Recursive adaptation in action: allochronic isolation and divergence of host-associated populations of the apple maggot fly,Rhagoletis pomonella,following its recent introduction to the western USA

An outstanding issue in the study of insect host races concerns the idea of ‘recursive adaptive divergence’, whereby adaptation can occur repeatedly across space and/or time, and the most recent adaptive episode is defined by one or more previously similar cases. The host plant shift of the apple maggot fly, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae, Carpomyini), from ancestral downy hawthorn [Crataegus mollis (Torr. & A. Gray) Scheele] to introduced, domesticated apple (Malus domestica Borkh.) in the eastern USA has long served as a model system for investigating ecologically driven host race formation in phytophagous insect specialists. Here, we report results from an annual geography survey of eclosion time demonstrating a similar ecological pattern among nascent host-associated populations of the fly recently introduced ca. 40 years ago from its native range in the east into the Pacific Northwest (PNW) region of the USA. Specifically, using data collected from 25 locations across 5 years, we show that apple-infesting fly populations in the PNW have rapidly and repeatedly shifted (and maintained differences in) their adult eclosion life-history timing to infest two novel hawthorn hosts with different fruiting phenologies – a native species (Crataegus douglasii Lindl.) and an introduced species (Crataegus monogyna Jacq.) – generating partial allochronic reproductive isolation in the process. The shifts in the PNW parallel the classic case of host race formation in the eastern USA, but have occurred bi-directionally to two hawthorn species with phenologies slightly earlier (black hawthorn) and significantly later (ornamental hawthorn) than apple. Our results imply that R. pomonella can both possess and retain extensive-standing variation (i.e., ‘adaptive memory’) in diapause traits, even following introductions, to rapidly and temporally track novel phenological host opportunities when they arise. Thus, ‘specialized’ host races may not constitute evolutionary dead ends. Rather, adaptive phenotypic and genetic memory may carry over from one host shift to the next, recursively facilitating host race formation in phytophagous insects.     

Colonization,stability, and adaptation in a transplant experiment of the polymorphic land snail Cepaea nemoralis (Gastropoda: Pulmonata) at the edge of its geographical range

At the eastern margins of the geographical distribution in Europe, populations of Cepaea nemoralis are sparse and limited to urban environments to which they are possibly confined by relatively warmer climates. In 1999 we introduced 1101 C. nemoralis individuals originating from nine urban populations to a rural location in the area. The snails established a viable population, which suggests that confinement to urban settings is dispersal‐ rather than climate‐limited. The snails filled available habitats at a rate of approximately 400–600 m² year^?1. On the whole, morph frequencies remained remarkably stable; changes that occurred are attributable to segregation of alleles or chromosomes. However, snails responded to habitat heterogeneity: consistent and predictable divergence occurred between habitat types, such that light‐shelled snails were repeatedly more frequent in the open than in adjoining shaded habitats. This suggests the operation of climatic and/or visual selection. As the whole area encompassing seven distinct habitat patches was only 0.3 ha, and the maximum duration of population divergence was only 11 years (fewer than four snail generations), these results indicate extremely small temporal and spatial scales of adaptation during initial phases of population establishment and spread. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 104 , 462–470.     

Factors influencing in vitro infectivity and growth of Rickettsia peacockii (Rickettsiales: Rickettsiaceae), an endosymbiont of the Rocky Mountain wood tick, Dermacentor andersoni (Acari, Ixodidae)

Rickettsia peacockii, a spotted fever group rickettsia, is a transovarially transmitted endosymbiont of Rocky Mountain wood ticks, Dermacentor andersoni. This rickettsia, formerly known as the East Side Agent and restricted to female ticks, was detected in a chronically infected embryonic cell line, DAE100, from D. andersoni. We examined infectivity, ability to induce cytopathic effect (CPE) and host cell specificity of R. peacockii using cultured arthropod and mammalian cells. Aposymbiotic DAE100 cells were obtained using oxytetracycline or incubation at 37 degrees C. Uninfected DAE100 sublines grew faster than the parent line, indicating R. peacockii regulation of host cell growth. Nevertheless, DAE100 cellular defenses exerted partial control over R. peacockii growth. Rickettsiae existed free in the cytosol of DAE100 cells or within autophagolysosomes. Exocytosed rickettsiae accumulated in the medium and were occasionally contained within host membranes. R. peacockii multiplied in other cell lines from the hard ticks D. andersoni, Dermacentor albipictus, Ixodes scapularis, and Ixodes ricinus; the soft tick Carios capensis; and the lepidopteran Trichoplusia ni. Lines from the tick Amblyomma americanum, the mosquito Aedes albopictus, and two mammalian cell lines were non-permissive to R. peacockii. High cell densities facilitated rickettsial spread within permissive cell cultures, and an inoculum of one infected to nine uninfected cells resulted in the greatest yield of infected tick cells. Cell-free R. peacockii also were infectious for tick cells and centrifugation onto cell layers enhanced infectivity approximately 100-fold. The ability of R. peacockii to cause mild CPE suggests that its pathogenicity is not completely muted. An analysis of R. peacockii-cell interactions in comparison to pathogenic rickettsiae will provide insights into host cell colonization mechanisms.     

Host range validation,molecular identification and release and establishment of a Chinese biotype of the Asian leaf beetle Lilioceris cheni (Coleoptera: Chrysomelidae: Criocerinae) for control of Dioscorea bulbifera L. in the southern United States

Dioscorea bulbifera, an Asian vine, is invasive in the southeastern USA. It rarely flowers but propagates from potato-like bulbils formed in leaf axils, which persist into the subsequent growing season. Lilioceris cheni Gressitt and Kimoto, a foliage-feeding beetle (Coleoptera: Chrysomelidae: Criocerinae) from Nepal, had been tested, proven to be a specialist and approved for release as a biological control agent. Regulatory delays, however, resulted in the demise of quarantine-held colonies, and acquisition of new Nepalese stock proved untenable. Searches then undertaken in southern China resulted in the collection of over 300 similar beetles. Two Chinese Lilioceris species were identified: one confirmed to be L. cheni and the other identified as Lilioceris egena (Weise). Mitochondrial analysis revealed an exact DNA match between some Chinese and one of the two Nepalese c oxidase subunit I haplotypes and all Chinese L. cheni haplotypes clustered as a single species but the comingling of the two species aroused concerns over possible hybridisation. These concerns were allayed by nuclear D2 analysis showing the absence of dual parental sequences. Nonetheless, diligence was exercised to ensure that the Chinese strains were safe to release. Abridged host testing using critical test species verified specificity. Caged releases during autumn 2011 documented the ability of adult beetles to overwinter in south Florida despite a prolonged lack of foliage. Open releases the following year produced vigorous populations that caused extensive defoliation. Preliminary observations indicate that L. cheni now contributes to the control of D. bulbifera and the bulbil-feeding L. egena should complement these effects if its host range proves appropriate.     

Relationships of song structure to phylogenetic history,habitat, and morphology in the vireos,greenlets, and allies (Passeriformes: Vireonidae)

Acoustic signals show immense variation among passerines, and several hypotheses have been proposed to explain this diversity. In this study, we tested, for the first time, the relationships of song structure to phylogeny, habitat type, and morphology in the vireos and allies (Vireonidae). Every measure of song structure considered in this study had moderate and significant phylogenetic signal. Furthermore, two song-constraining morphological traits, bill shape and body mass, also exhibited significant phylogenetic signal. Song length showed the largest within-clade similarity; longer songs were highly conserved in part of the greenlet (Hylophilus) clade, whereas shorter songs characterized the remaining seven genera. We found no differences in song structure among vireonids living in different habitat types. However, vireonids with shorter, stouter bills and larger bodies sang songs with lower minimum and maximum peak frequency, compared with species with longer, thinner bills and smaller bodies. We conclude that Vireonidae song evolution is driven partially by phylogenetically conserved morphological traits. Our findings support the phylogenetic signal and morphological constraints hypotheses explaining structural diversity in avian acoustic signals.     

Morphometric and molecular variation in concert: taxonomy and genetics of the reticulate Pyrenean and Iberian alpine spiny fescues (Festuca eskia complex,Poaceae)

The Iberian mountain spiny fescues are a reticulate group of five diploid grass taxa consisting of three parental species and two putative hybrids: F. × souliei (F. eskia × F. quadriflora) and F. × picoeuropeana (F. eskia × F. gautieri). Phenotypic and molecular studies were conducted with the aim of determining the taxonomic boundaries and genetic relationships of the five taxa and disentangling the origins of the two hybrids. Statistical analyses of 31 selected phenotypic traits were conducted on individuals from 159 populations and on nine type specimens. Molecular analyses of random amplified polymorphic DNA (RAPD) markers were performed on 29 populations. The phenotypic analyses detected significant differences between the five taxa and demonstrated the overall intermediacy of the F. × picoeuropeana and F. × souliei between their respective parents. The RAPD analysis corroborated the genetic differentiation of F. eskia, F. gautieri and F. quadriflora and the intermediate nature of the two hybrids; however, they also detected genetic variation within F. × picoeuropeana. These results suggest distinct origins for F. × picoeuropeana in the Cantabrian and Pyrenean mountains, with the sporadic Pyrenean populations having potentially resulted from recent hybridizations and the stabilized Cantabrian ones from older events followed by potential displacements of the parents. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 676–706.     

Developmental genetic profiles of glutamate receptor system,neuromodulator system,protector of normal tissue and mitochondria,and reelin in marmoset cortex: Potential molecular mechanisms of pruning phase of spines in primate synaptic formation process during the end of infancy and prepuberty (II)

This is the second report of a series paper, which reports molecular mechanisms underlying the occurrence of pruning spine phase after rapid spinogenesis phase in neonates and young infant in the primate brain. We performed microarray analysis between the peak of spine numbers [postnatal 3 months (M)] and spine pruning (postnatal 6 M) in prefrontal, inferior temporal, and primary visual cortices of the common marmoset (Callithrix jacchus). The pruning phase is not clearly defined in rodents but is in primates including the marmoset. The differentially expressed genes between 3 M and 6 M in all three cortical areas were selected by two-way analysis of variance. The list of selected genes was analyzed by canonical pathway analysis using “Ingenuity Pathway Analysis of complex omics data” (IPA; Ingenuity Systems, Qiagen, Hilden, Germany). In this report, we discuss these lists of genes for the glutamate receptor system, G-protein-coupled neuromodulator system, protector of normal tissue and mitochondria, and reelin. (1) Glutamate is a common neurotransmitter. Its receptors AMPA1, GRIK1, and their scaffold protein DLG4 decreased as spine numbers decreased. Instead, GRIN3 (NMDA receptor) increased, suggesting that strong NMDA excitatory currents may be required for a single neuron to receive sufficient net synaptic activity in order to compensate for the decrease in synapse. (2) Most of the G protein-coupled receptor genes (e.g., ADRA1D, HTR2A, HTR4, and DRD1) in the selected list were upregulated at 6 M. The downstream gene ROCK2 in these receptor systems plays a role of decreasing synapses, and ROCK2 decreased at 6 M. (3) Synaptic phagosytosis by microglia with complement and other cytokines could cause damage to normal tissue and mitochondria. SOD1, XIAP, CD46, and CD55, which play protective roles in normal tissue and mitochondria, showed higher expression at 6 M than at 3 M, suggesting that normal brain tissue is more protected at 6 M. (4) Reelin has an important role in cortical layer formation. In addition, RELN and three different pathways of reelin were expressed at 6 M, suggesting that new synapse formation decreased at that age. Moreover, if new synapses were formed, their positions were free and probably dependent on activity.     

The centipede genus Eupolybothrus Verhoeff, 1907 (Chilopoda: Lithobiomorpha: Lithobiidae) in North Africa, a cybertaxonomic revision, with a key to all species in the genus and the first use of DNA barcoding for the group

The centipede genus Eupolybothrus Verhoeff, 1907 in North Africa is revised. A new cavernicolous species, Eupolybothruskahfi Stoev & Akkari, sp. n., is described from a cave in Jebel Zaghouan, northeast Tunisia. Morphologically, it is most closely related to Eupolybothrusnudicornis (Gervais, 1837) from North Africa and Southwest Europe but can be readily distinguished by the long antennae and leg-pair 15, a conical dorso-median protuberance emerging from the posterior part of prefemur 15, and the shape of the male first genital sternite. Molecular sequence data from the cytochrome c oxidase I gene (mtDNA-5' COI-barcoding fragment) exhibit 19.19% divergence between Eupolybothruskahfi and Eupolybothrusnudicornis, an interspecific value comparable to those observed among four other species of Eupolybothrus which, combined with a low intraspecific divergence (0.3-1.14%), supports the morphological diagnosis of Eupolybothruskahfi as a separate species. This is the first troglomorphic myriapod to be found in Tunisia, and the second troglomorph lithobiomorph centipede known from North Africa. Eupolybothrusnudicornis is redescribed based on abundant material from Tunisia and its post-embryonic development, distribution and habitat preferences recorded. Eupolybothruscloudsley-thompsoni Turk, 1955, a nominal species based on Tunisian type material, is placed in synonymy with Eupolybothrusnudicornis. To comply with the latest technological developments in publishing of biological information, the paper implements new approaches in cybertaxonomy, such as fine granularity XML tagging validated against the NLM DTD TaxPub for PubMedCentral and dissemination in XML to various aggregators (GBIF, EOL, Wikipedia), vizualisation of all taxa mentioned in the text via the dynamically created Pensoft Taxon Profile (PTP) page, data publishing, georeferencing of all localities via Google Earth, and ZooBank, GenBank and MorphBank registration of datasets. An interactive key to all valid species of Eupolybothrus is made with DELTA software.     

<Emphasis Type="Italic">Kuth</Emphasis> (<Emphasis Type="Italic">Saussurea lappa</Emphasis>) cultivation in the cold desert environment of the Lahaul valley,northwestern Himalaya,India: arising threats and need to revive socio-economic values

Surveys were conducted in the cold desert environment of the Lahaul valley in the northwestern Himalaya for assessing the past and present status of Kuth (Saussurea lappa) cultivation. The findings reveal that this age-old practice now is in bottleneck. Main factors responsible for this setback to the species were the lengthy cultivation cycle, small land holdings, and even fluctuating and relatively low market prices. Owing to these constraints farmers have now started replacing cultivation of this threatened herb with pea (Pisum sativum L.), potato (Solanum tuberosum L.) and hop (Humulus lupulus L.). These crops obtained popularity due to comparatively more economic returns as well as their easy adaptability to the short growth season of the cold desert environment. Kuth cultivation in this region is among the interesting examples of domesticating wild medicinal herb by some innovative farmers during the 1920s. However, in the recent past farmers have been less interested to continue this practice due to its larger cultivation cycle, more profits with cash crops like pea and potato, and permit formalities at the time of export from the valley. In addition to being the oldest cash crop in the cold desert environment, Kuth is an endangered medicinal herb that has to be conserved on a priority basis. This study attempts to find out potential measures such as regular revision of market rates, development of existing uncultivable land under medicinal plant cultivation and strengthening the marketing network through establishment of federations of farmers at village level to revive cultivation of this important species.     

Distribution of Elements in the Millipede, <Emphasis Type="BoldItalic">Oxidus gracilis</Emphasis> C. L. Koch (Polydesmida: Paradoxosomatidae) and the Relation to Environmental Habitats

The concentrations of 55 elements in the millipede, Oxidus gracilis, soil and plant in the habitat were examined using inductively coupled plasma atomic emission spectrometry (ICP-AES) and inductively coupled plasma mass spectrometry (ICP-MS). In all the millipedes, Ca concentration derived from calcium carbonate in the exoskeleton was the highest at average 94 μ/mg-weight. The other major elements were the following: Mg, K, Na, Zn, Fe, Al, Cu, Sr, Ba, Mn and Ti (> 1 ng/mg-body weight), whereas Se, Mo, Ag, Cd, Co, Li and Ce etc. were in trace levels. Interestingly, the various 15 elements such as Ca, Na, Zn, Al, Ba, Ga, Ag, Cd, Co and Y in environmental habitats were well reflected in the body of the millipede. Although the heavy metal contents, in the order of Cu>Pb>Cd, were similar to those of other invertebrates, Cu in the millipede was remarkably high concentration. Zn was maintained in a range of 72–394 ng/mg-weight as essential element in the body and no difference was found in the sexes. The C1 chondrite normalization pattern for lanthanoid series elements in the millipede, soil and plant indicated that the environmental habitats were well protected from pollution. These characteristics of internal elements and metal accumulation in the millipede or relation to their habitats would be useful information for the environmental pollution studies.